LOCUS       sequence-152440-        6023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of N-terminal fusion of mClover3 to Tubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6023)
  AUTHORS   Bajar BT, Wang ES, Lam AJ, Kim BB, Jacobs CL, Howe ES, Davidson MW, 
            Lin MZ, Chu J
  TITLE     Improving brightness and photostability of green and red fluorescent
            proteins for live cell imaging and FRET reporting.
  JOURNAL   Sci Rep. 2016 Feb 16;6:20889. doi: 10.1038/srep20889.
  PUBMED    26879144
REFERENCE   2  (bases 1 to 6023)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2016-02-16"; volume: "6:20889. doi"; pages: " 
            10.1038/srep20889"
COMMENT     Sequence Label: pKanCMV-mClover3-18aa-Tubulin
FEATURES             Location/Qualifiers
     source          1..6023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2869..2990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2906..2925)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2960..2979
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2997..3452)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3134..3155)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3346..3365
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3479..3583
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3585..3942
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3585..3776
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3608..3628)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3793..3928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3977..4771
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3977..4771
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4031..4050)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4641..4660
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4959..4978)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5003..5050
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5379..5967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5868..5887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgtc taagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga gggcgagggc
      721 gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccttcggc tacggcgtgg cctgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct ctttcaagga cgacggtacc tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaacttc aacagccact acgtctatat cacggccgac
     1081 aagcagaaga actgcatcaa ggctaacttc aagatccgcc acaacgttga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag ccatcagtcc aagctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga ttacacatgg catggacgag
     1321 ctgtacaagt ccggactcag atctggcagc ggtggaggca gcgcatccgg cggaagcgga
     1381 agcgtgcgtg agtgcatctc catccacgtt ggccaggctg gtgtccagat tggcaatgcc
     1441 tgctgggagc tctactgcct ggaacacggc atccagcccg atggccagat gccaagtgac
     1501 aagaccattg ggggaggaga tgactccttc aacaccttct tcagtgagac gggcgctggc
     1561 aagcacgtgc cccgggctgt gtttgtagac ttggaaccca cagtcattga tgaagttcgc
     1621 actggcacct accgccagct cttccaccct gagcagctca tcacaggcaa ggaagatgct
     1681 gccaataact atgcccgagg gcactacacc attggcaagg agatcattga ccttgtgttg
     1741 gaccgaattc gcaagctggc tgaccagtgc accggtcttc agggcttctt ggttttccac
     1801 agctttggtg ggggaactgg ttctgggttc acctccctgc tcatggaacg tctctcagtt
     1861 gattatggca agaagtccaa gctggagttc tccatttacc cagcacccca ggtttccaca
     1921 gctgtagttg agccctacaa ctccatcctc accacccaca ccaccctgga gcactctgat
     1981 tgtgccttca tggtagacaa tgaggccatc tatgacatct gtcgtagaaa cctcgatatc
     2041 gagcgcccaa cctacactaa ccttaaccgc cttattagcc agattgtgtc ctccatcact
     2101 gcttccctga gatttgatgg agccctgaat gttgacctga cagaattcca gaccaacctg
     2161 gtgccctacc cccgcatcca cttccctctg gccacatatg cccctgtcat ctctgctgag
     2221 aaagcctacc atgaacagct ttctgtagca gagatcacca atgcttgctt tgagccagcc
     2281 aaccagatgg tgaaatgtga ccctcgccat ggtaaataca tggcttgctg cctgttgtac
     2341 cgtggtgacg tggttcccaa agatgtcaat gctgccattg ccaccatcaa aaccaagcgc
     2401 agcatccagt ttgtggattg gtgccccact ggcttcaagg ttggcatcaa ctaccagcct
     2461 cccactgtgg tgcctggtgg agacctggcc aaggtacaga gagctgtgtg catgctgagc
     2521 aacaccacag ccattgctga ggcctgggct cgcctggacc acaagtttga cctgatgtat
     2581 gccaagcgtg cctttgttca ctggtacgtg ggtgagggga tggaggaagg cgagttttca
     2641 gaggcccgtg aagatatggc tgcccttgag aaggattatg aggaggttgg tgtggattct
     2701 gttgaaggag agggtgagga agaaggagag gaatactaag gatccaccgg atctagataa
     2761 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2821 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     3001 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3061 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3121 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3181 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3241 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3301 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3361 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3421 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3481 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3541 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3601 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3661 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3721 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3781 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3841 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3901 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3961 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4021 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4081 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4141 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4201 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4261 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4321 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4381 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4441 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4501 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4561 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4621 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4681 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4741 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4801 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4861 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4921 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4981 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5041 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5101 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5161 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5221 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5281 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5341 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5401 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5461 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5521 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5581 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5641 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5701 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5761 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5821 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5881 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5941 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6001 gccttttgct ggccttttgc tca
//
